Publication Database

Measuring Ca(2+) binding to short chain fatty acids and gluconate with a Ca(2+) electrode:
role of the reference electrode (2014)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Stumpff, Friederike (WE 2)

McGuigan, J.A. (WE 2)

Quelle

Analytical biochemistry

Bandzählung: 459

Seiten: 46 – 52

ISSN: 0003-2697

Sprache

Englisch

Verweise

DOI: 10.1016/j.ab.2014.05.001

Pubmed: 24835426

Kontakt

Institut für Veterinär-Physiologie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62600
physiologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

Many organic anions bind free Ca2+, the total concentration of which must be adjusted in experimental solutions. Because published values for the apparent dissociation constant (Kapp) describing the Ca2+ affinity of short chain fatty acids (SCFAs) and gluconate are highly variable, Ca2+ electrodes coupled to either a 3M KCl or Na+ selective electrode were used to redetermine Kapp. All solutions contained 130mM Na+, whereas the concentration of the studied anion was varied from 15 to 120mM, replacing Cl- that was decreased concomitantly to maintain osmolarity. This induces changes in the liquid junction potential (LJP) at the 3M KCl reference electrode, leading to a systematic underestimation of Kapp if left uncorrected. Because the Na+ concentration in all solutions was constant, an Na+ electrode was used to directly measure the changes in the LJP at the 3 M KCl reference, which were small but twice those predicted by the Henderson equation. Determination of Kapp either after correction for these LJP changes or via direct reference to an Na+ electrode showed that SCFAs do not bind Ca2+ and that the Kapp for the binding of Ca2+ to gluconate at pH 7.4, ionic strength 0.15M, and 23°C was 52.7mM.